Whole Saliva Of Patients Research Articles

In vivo analysis of secreted aspartyl proteinase expression in human oral candidiasis.

Secreted aspartyl proteinases are putative virulence factors in Candida infections. Candida albicans possesses at least nine members of a SAP gene family, all of which have been sequenced. Although the expression of the SAP genes has been extensively characterized under laboratory growth conditions, no studies have analyzed in detail the in vivo expression of these proteinases in human oral colonization and infection. We have developed a reliable and sensitive procedure to detect C. albicans mRNA from whole saliva of patients with oral C. albicans infection and those with asymptomatic Candida carriage. The reverse transcription-PCR protocol was used to determine which of the SAP1 to SAP7 genes are expressed by C. albicans during colonization and infection of the oral cavity. SAP2 and the SAP4 to SAP6 subfamily were the predominant proteinase genes expressed in the oral cavities of both Candida carriers and patients with oral candidiasis; SAP4, SAP5, or SAP6 mRNA was detected in all subjects. SAP1 and SAP3 transcripts were observed only in patients with oral candidiasis. SAP7 mRNA expression, which has never been demonstrated under laboratory conditions, was detected in several of the patient samples. All seven SAP genes were simultaneously expressed in some patients with oral candidiasis. This is the first detailed study showing that the SAP gene family is expressed by C. albicans during colonization and infection in humans and that C. albicans infection is associated with the differential expression of individual SAP genes which may be involved in the pathogenesis of oral candidiasis.

Interindividual differences in degradation of sodium monofluorophosphate by saliva in relation to oral health status

The enzymatic degradation of sodium monofluorophosphate by whole saliva in patients with differing oral health status was compared. Ten patients each with good or poor oral health were selected. Assessment of oral health status included indices of the amount of dental plaque, caries experience and counts of Streptococcus mutans and lactobacilli. Whole-saliva samples were collected under standardized conditions and monofluorophosphatase (MFPase) activity was measured in 2-ml saliva samples during 2 h of incubation at 37°C with 1 ml of monofluorophosphate solution (equivalent 31.5 mmol/1 F −). MFPase activity was found in all the saliva samples. The values ranged from 0.6 to 17.8 nmol F −/ml saliva per min. High MFPase activities correlated well with large amounts of plaque, with the D component of the DMFT index and with high counts of salivary S. mutans and lactobacilli. The results show a wide range of MFPase activity in individuals and a statistically significant correlation ( p < 0.05) between high MFPase activity and poor oral health status.

Salivary peroxidase activity in whole saliva of patients with insulin-dependent (type-1) diabetes mellitus.

In this study, salivary peroxidase activity was measured in a group of 10 patients with insulin-dependent (type I) diabetes mellitus (IDDM) who had a tendency towards periodontitis. In healthy subjects (N : 10), mean salivary peroxidase activity was 0.0025 +/- 0.001 IU/ml, while in this group of type I diabetic patients it was 0.051 +/- 0.015 IU/ml, a significantly higher level (p < 0.001). Approximal plaque index (API), modified sulcus bleeding index (mod SBI) and pocket depths were assessed clinically. The values for mod SBI and API were 60% and 68% respectively for the diabetic patients while for the control group mod SBI was measured 0.0% and the value for API was 10.67% (p < 0.001). The administration of this simple and practical test may provide an early marker of a tendency towards periodontitis in IDDM patients.

Peroxidases, lactoferrin and lysozyme in peripheral blood neutrophils, gingival crevicular fluid and whole saliva of patients with localized juvenile periodontitis.

The aim of this study was to examine the longitudinal association of selected non-immune anti-microbial host factors (peroxidases, lysozyme and lactoferrin) to the localized juvenile periodontitis (LJP) disease status. Peroxidases, lysozyme and lactoferrin were quantitated from seven patients with LJP before and after periodontal therapy. Analyses were performed from simultaneously collected samples of peripheral blood polymorphonuclear leukocytes (PMNs), gingival crevicular fluid (GCF from diseased sites) and paraffin-stimulated whole saliva. Similar assays were done also from seven periodontally healthy controls. During untreated phase of LJP myeloperoxidase, lysozyme and lactoferrin concentrations were remarkably elevated in peripheral blood PMNs, also reflected in their high concentrations in GCF. All these values normalised with respect to healthy controls during the periodontal therapy. No similar longitudinal changes were seen in whole saliva but during therapy salivary peroxidase concentrations declined below the control values, in accordance with our previous observations in parotid saliva samples of LJP patients. In LJP the concentrations of lysozyme, lactoferrin and myeloperoxidase are significantly elevated in peripheral blood PMNs, also reflected in GCF. During periodontal therapy these values decline and approach those observed in healthy controls. No similar changes are seen in stimulated whole saliva.

Myeloperoxidase activity in peripheral blood, neutrophil crevicular fluid and whole saliva of patients with periodontal disease.

Mean levels of myeloperoxidase (MPO) activity were determined in samples of gingival crevicular fluid (GCF), whole saliva and peripheral blood neutrophils from patients with rapidly progressive periodontitis (RPP) and adult periodontitis (AP) using a spectrophotometric method. The mean neutrophil MPO activity in the RPP group was 563.1 +/- 137.05 U/l x 10(6)/ml, that in the AP group was 483.3 +/- 88.81 U/l x 10(6)/ml, and that in the control group was 220.6 +/- 26.7 U/l x 10(6)/ml. The mean GCF MPO activity in the RPP group was 15.13 +/- 2.34 U/mg, which was significantly higher than in the other two groups. The mean whole saliva MPO activity in the RPP group was 0.14 +/- 0.04 U/ml, that in the AP group was 0.11 +/- 0.02 U/ml, and that in the control group was 0.05 +/- 0.06 U/ml. MPO activity detected in the samples was significantly increased in the patient groups when compared to the healthy subjects. The highest MPO activity was found in the RPP group. The present findings suggest a relationship between MPO activity and the pattern and severity of periodontal breakdown. Also the increased MPO activity in periodontally diseased patients can be attributed to the increased number of neutrophils, the degranulation of these cells and also their hyperactive state in the presence of chronic antigenic stimulation.

Qualitative and quantitative studies of Gc (vitamin D-binding protein) in normal subjects and patients with periodontal disease.

Qualitative and quantitative studies of Gc (Vitamin D‐binding protein) were carried out with whole and parotid saliva from 13 patients with periodontal disease, and 19 control subjects. Using an ELISA technique, a low concentration of Gc was found in the parotid saliva of all subjects. The whole saliva of patients with periodontal disease contained concentrations of Gc which were significantly increased over those in either dentulous or edentulous control subjects (p&lt;0.01). In addition, the Gingival Index scores of these patients correlated with the concentration of Gc (r = 0.87; p&lt;0.01). Rocket immunoelectrophoresis showed that &gt; 90% of Gc in saliva from these patients was complexed with G‐actin.

Immunoglobulins and innate antimicrobial factors in whole saliva of patients with insulin-dependent diabetes mellitus.

We analyzed the flow rate and composition of paraffin-stimulated whole saliva samples from 35 adult diabetic patients and their age- and sex-matched, non-diabetic, clinically healthy controls. All patients had insulin-dependent diabetes (IDDM) with a mean (+/- S.D.) duration of 14.0 +/- 9.1 years. The saliva analysis included the quantitation of total protein, amylase, immunoglobulins (isotypes A, G, and M), and the non-antibody, innate antimicrobial factors (lysozyme, lactoferrin, salivary peroxidase, myeloperoxidase, thiocyanate, and hypothiocyanite). The whole saliva samples from diabetic patients had significantly higher amounts of IgA (p less than 0.001) and IgG (p less than 0.05) than did the controls. No differences between the study groups were observed in flow rate, protein content, amylase activity, or IgM. The levels of innate defense factors were similar in both study groups except for salivary peroxidase, which was higher (p less than 0.02) among diabetics than among controls. Our results indicate that the antimicrobial defense capacity of whole saliva is not impaired in diabetic patients.

Adrenal status assessed by direct radioimmunoassay of cortisol in whole saliva or parotid saliva.

We describe a direct radioimmunoassay for cortisol in 10-microliter volumes of parotid saliva or whole saliva. Binding proteins are absent from these fluids, as demonstrated by the excellent correlation between results for samples assayed directly and by a comparison procedure involving extraction with 1,2-dichloroethane. The direct assay is specific, precise, and had a lower limit of sensitivity of 4 pg per assay tube. Comparison of cortisol concentrations in plasma, parotid saliva, and whole saliva in persons undergoing investigations for assessing adrenal function, including stimulation with cosyntropin (Synacthen) and suppression with dexamethasone, indicated that changes in plasma cortisol concentration were accurately and immediately reflected in saliva from either the parotidgland or whole saliva. A marked circadian rhythm has also been demonstrated for cortisol in parotid-gland saliva and whole saliva. We had to modify the 1,2-dichloroethane extraction procedure for accurate determination of cortisol in parotid saliva and whole saliva of patients undergoing treatment with metyrapone.

The association of the IGA levels of serum and whole saliva with the progression of oral cancer.

Elevated IgA concentrations in the serum and whole saliva of patients with neoplastic disease have been reported. However, there does not seem to be any information available on the correlation of the secretory immunoglobulins of these patients with the progress of their disease. The whole saliva and sera of 102 cases of squamous carcinoma of the oral mucosa were studied. Patients were subclassified into age-matched groups of primary untreated cancer (21), recurrent cancer (18), and "cured" patients who had been free of disease for at least 9 months (16). A separate group of laryngeal (14) patients was included. Sixteen (16) patients treated by radiation therapy alone were also included to test the effects of radiation on whole saliva IgA. An age-matched group of patients (17) with moderate to severe periodontal disease served as controls. Unstimulated whole saliva was collected for 15 minutes and stored at 0 degrees C. IgA was quantitated by radial immunodiffusion on Hyland and Meloy Laboratory low level plates with 7S and 11S standards. Serum IgA was measured on Hyland plates with serum standards. Protein content of saliva was assayed by the Lowry method. Flow rate was quantitated for the 15-minute collection period. Primary oral and laryngeal cancer patients had a two-fold increase of serum and salivary IgA compared to controls. Recurrent cancer patients had even greater elevation of salivary IgA. Cured patients showed a persistent elevation of serum but a return to normal of salivary IgA. Radiation therapy did not markedly influence the level of salivary IgA. A followup study of 26 patients confirmed this pattern of a drop in whole salivary IgA with cure and a spike with recurrence. It would appear that IgA levels of whole saliva and serum are elevated in oral cancer patients, and that salivary IgA levels may prove useful in distinguishing patients with possible recurrent disease.

Salivary IgA in periodontal disease.

The IgA content in whole and parotid saliva was determined in normal human subjects and in individuals afflicted with periodontal disease. The immunological measurements were made by means of a diffusion-in-gel method previously described. Elevated IgA-levels were found in the whole saliva of patients with periodontal disease, while normal levels of IgA were noted in sera and parotid saliva of all subjects studied regardless of their periodontal status. It was concluded that the elevated levels of IgA in the whole saliva was due to increased leakage of IgA via the gingival fluid.